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1.
Braz. j. biol ; 79(2): 169-173, Apr.-June 2019. tab, graf
Article in English | LILACS | ID: biblio-989440

ABSTRACT

Abstract Even today, an effective diagnostic test for aspergillosis in penguins is unknown, being the gold standard post-mortem examinations. The fungal antigen galactomannan (GM) has been used as a biomarker of disease in humans and is detected by the Platelia Aspergillus EIA (BioRad)®, a commercial kit based on the sandwich ELISA technique. It is standardized for use in neutropenic patients, however studies have demonstrated its usefulness also possible for birds. The aim of our study was to evaluate the effectiveness of Platelia Aspergillus EIA® test (BioRad-US) in the diagnosis of aspergillosis in Magellanic penguins, determining sensitivity, specificity, and positive and negative predictive values for different cut-off points. Were included in the study, blood serum samples (n = 29) Magellanic penguins in captivity that died by aspergillosis. Detection of GM was performed following manufacturer's instructions and the GM index was obtained by dividing the average value of OD of the duplicate of the clinical sample by duplicate OD of the average value of the cut-off sample provided by the kit. Through information database results were obtained for the presence of anti-Aspergillus fumigatus antibodies detected by agar gel immunodiffusion (AGID) for all serum samples. Results were analyzed using chi-square test and Kruskal-Wallis from SPSS 20.0, IBM®. ROC curve was obtained and from this, rates of sensitivity, specificity, positive and negative predictive values were also calculated based on four different cutoff points (0.5, 1.0, 1.5 and 2.0). The serum GM index did not differ between animals of the case and control group (pkw =0.097). In determining the ROC curve for serum GM detection the value of area under the curve was 0.635. From the values ​​determined by the coordinate of the curve, four different cut points (0.5, 1.0, 1.5 and 2.0) were analyzed, resulting in sensitivity rates ranging from 86.2 to 34.5% % and specificity between 87% and 26.1%. By comparing the serum GM index in group case as the presence or absence of antibodies detected by AGID was found p=0.503. The detection of GM the Platelia Aspergillus EIA® test seems is not be useful for the diagnosis of aspergillosis in naturally infected penguins.


Resumo Ainda hoje, um teste diagnóstico eficaz para aspergilose em pinguins não é conhecido, sendo o padrão-ouro os exames post-mortem. O antígeno fúngico galactomanana (GM) tem sido utilizado como biomarcador da doença em humanos, sendo detectado pelo Platelia Aspergillus EIA (BioRad)®, um kit comercial que se baseia na técnica ELISA sanduíche. É padronizado para utilização em pacientes neutropênicos, no entanto estudos tem demonstrado sua possível utilidade também para aves.O objetivo de nosso estudo foi avaliar a eficácia do teste Platelia Aspergillus EIA® (BioRad-US) no diagnóstico da aspergilose em pinguins-de-Magalhães, determinando sensibilidade, especificidade e valores preditivos positivos e negativos em diferentes pontos de corte. Foram incluídas no estudo, amostras de soro sanguíneo (n=29) de pinguins-de-Magalhães em cativeiro que vieram a óbito por aspergilose. A detecção de GM foi realizada seguindo instruções do fabricante e o índice de GM foi obtido dividindo o valor da média da DO da duplicata da amostra clínica pelo valor da média da DO da duplicata da amostra de cut-off fornecida pelo kit. Através de informações em banco de dados foram obtidos resultados sobre a presença de anticorpos anti-Aspergillus fumigatus, detectada por Imunodifusão em gel de ágar (IDGA) em todas as amostras séricas. Os resultados foram analisados utilizando-se teste de qui-quadrado e Kruskal-Wallis a partir do programa estatístico SPSS 20.0, IBM®. Curva ROC foi obtida e a partir desta, taxas de sensibilidade, especificidade, valores preditivo positivo e negativo foram igualmente calculados considerando quatro diferentes pontos de corte (0.5, 1.0, 1.5 e 2.0). O índice de GM sérica não diferiu entre os animais do grupo caso e controle (pKW = 0.097). Na determinação da curva ROC para detecção de GM sérica o valor da área sobre a curva foi de 0.635. A partir dos valores determinados pelas coordenadas da curva, quatro diferentes pontos de corte (0.5, 1.0, 1.5 e 2.0) foram analisados, resultando em taxas de sensibilidade variando de 86.2% a 34.5%, e de especificidade entre 87% e 26.1%. Ao comparar o índice de GM sérica nos animais do grupo caso quanto a presença ou não de anticorpos detectados pela IDGA foi encontrado p=0.503. A detecção de GM pelo teste Platelia Aspergillus EIA® não parece ser útil para o diagnóstico da aspergilose em pinguins naturalmente infectados.


Subject(s)
Animals , Aspergillosis/diagnosis , Aspergillosis/veterinary , Aspergillus/isolation & purification , Enzyme-Linked Immunosorbent Assay/methods , Spheniscidae/microbiology , Biomarkers/analysis , Mannans/analysis
2.
Braz. j. med. biol. res ; 52(12): e8658, 2019. tab, graf
Article in English | LILACS | ID: biblio-1055464

ABSTRACT

Acute coronary syndromes are associated with a high prevalence of complications including heart failure (HF). The aim of this study was to investigate the association of novel biomarkers with the occurrence of post-acute myocardial infarction (AMI) HF. A prospective study was conducted with patients admitted to the emergency department with ST-segment elevation myocardial infarction (STEMI). Blood and urine samples were collected for analysis of traditional and novel biomarkers, including interleukin-6, vascular cell adhesion molecule 1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1). We compared the levels of these biomarkers between patients with and without post-STEMI HF. A total of 48 patients were assessed, with a prevalence of males. Fifteen patients (31.2%) had post-STEMI HF. Patients with HF had higher mean values of IL-6, VCAM-1, and ICAM-1 compared to those who did not develop HF (57.06 vs 14.03 pg/mL, P=0.001; 1719.58 vs 1304.34 ng/mL, P=0.001; and 1594.20 vs 1158.74 ng/mL, P<0.001, respectively). The three biomarkers were shown to be good predictors of post-STEMI HF (IL-6: AUC 0.786, P=0.002; VCAM-1: AUC 0.797, P=0.001; and ICAM-1: AUC 0.825, P<0.0001), with the respective cutoff points being calculated based on the best sensitivity and specificity indexes (IL-6: 8.67 pg/mL; VCAM-1: 1501.42 ng/mL; and ICAM-1: 1262.38 ng/mL). Of the three biomarkers, only VCAM-1 and ICAM-1 had a direct linear association between them (r=0.470, P<0.0001). IL-6, VCAM-1, and ICAM-1 were associated with the development of new post-AMI HF symptoms, but only VCAM-1 and ICAM-1 correlated with each other, possibly because they have the same pathophysiological mechanism of action.


Subject(s)
Humans , Male , Female , Aged , Interleukin-6/blood , Intercellular Adhesion Molecule-1/blood , Vascular Cell Adhesion Molecule-1/blood , Heart Failure/blood , Myocardial Infarction/blood , Biomarkers/blood , Case-Control Studies , Predictive Value of Tests , Prospective Studies , Sensitivity and Specificity
3.
Arq. bras. med. vet. zootec ; 67(1): 125-130, 2/2015. graf
Article in English | LILACS | ID: lil-741100

ABSTRACT

During the austral winter the appearance of the Magellanic penguin (Spheniscus magellanicus) is common on the southern coast of Brazil. The oil pollution constitutes a major cause of death of these birds. The monitoring of the weight and blood variables is important to make decisions during the rehabilitation and release of these animals. Thus, the aim of this study was to evaluate the relationship between the penguins' survival during rehabilitation and the values of hematocrit (Hct), total plasma protein (TPP) and body weight (BW) of the birds received at Centro de Recuperação de Animais Marinhos (Recovery Centre for Sea Animals) between 2006 and 2009. The analysis took place weekly, and penguins were classified according to the outcome of the rehabilitation process, when 101 specimens were rehabilitated and released, and 67 died. Released animals showed a decrease in the Hct in the second and third weeks (41±6% and 40±5% respectively) followed by recovery of the values in the fourth and fifth week (41±4% and 43±4% respectively). TPP levels increased steadily over the collections, stabilizing at 7.3±0.8g/dL in the fourth week. On the other hand, animals that died, although following the same response for TPP, showed no significant difference in Hct between collections, and showed a tendency for loss of BW in the last weeks of rehabilitation. The curves of BW and Hct during rehabilitation were considered a potential prognostic indicator of penguins in rehabilitation. Animals that arrived at the recovery centre with BW, Hct and TPP above 2.700g, 45% and 5.6 g / dL, respectively, have a greater chance of recovery, making these parameters crucial points in decision making regarding the treatment to be conducted during rehabilitation.


Durante o inverno austral, é comum o aparecimento do pinguim-de-Magalhães (Spheniscus magellanicus) no litoral sul do Brasil. A poluição por petróleo constitui a maior causa de morte dessas aves. O acompanhamento do peso e de variáveis sanguíneas é importante para a tomada de decisões durante a reabilitação e liberação desses animais. O objetivo deste estudo foi avaliar a relação entre a sobrevivência dos pinguins durante a reabilitação e os valores de hematócrito (Ht), proteínas plasmáticas totais (PPT) e peso corpóreo (PC) das aves recebidas no Centro de Recuperação de Animais Marinhos entre 2006 e 2009. As análises ocorreram semanalmente, e os pinguins foram classificados em função do desfecho do processo de reabilitação, quando 101 espécimes foram reabilitados e liberados e 67 foram a óbito. Animais liberados apresentaram queda no Ht na segunda e terceira semanas (41±6% e 40±5%, respectivamente), seguida de recuperação dos valores na quarta e quinta semanas (41±4% e 43±4%, respectivamente). Os níveis de PPT aumentaram progressivamente ao longo das coletas, estabilizando na quarta semana em 7,3±0,8g/dL. Em contrapartida, animais que foram a óbito, apesar de seguirem a mesma resposta para as PPT, não apresentaram diferença significativa do Ht, entre as coletas, e demonstraram tendência à perda de PC nas últimas semanas de reabilitação. As curvas de PC e Ht foram consideradas indicadoras potenciais de prognóstico durante a reabilitação dos pinguins. Animais que chegaram ao centro de recuperação com PC, Ht e PPT acima de 2.700g, 45% e 5,6g/dL, respectivamente, apresentaram maior chance de recuperação, fazendo desses parâmetros pontos críticos na tomada de decisão quanto ao tratamento a ser conduzido durante a reabilitação.


Subject(s)
Animals , Blood Proteins/analysis , Spheniscidae/blood
4.
Arq. bras. med. vet. zootec ; 66(6): 1703-1710, 12/2014. tab, graf
Article in English | LILACS | ID: lil-735782

ABSTRACT

The aim of this study was to evaluate somatic cell count (SCC), prevalence and etiology of mastitis in a dairy buffalo herd from Analândia, São Paulo State, Brazil, in the dry and rainy seasons. Additionally, antimicrobial susceptibility profile of microorganisms isolated from milk samples was also evaluated. 1,042 milk samples from female Murrah buffaloes in a dairy farm located in Analândia, São Paulo State, Brazil, collected between May 2011 and November 2012 were analyzed. After the mammary gland physical examination, strip cup test and California Mastitis Test (CMT) were performed. Afterwards, 50mL of milk samples from each mammary quarter were collected aseptically for SCC in automatic equipment and microbiological examination. The antimicrobial sensitivity profile to ampicillin, cefoperazone, ceftiofur, enrofloxacin, gentamicin, neomycin, oxacillin, penicillin, and sulfamethoxazole/trimethoprim was evaluated by disk diffusion method. The monthly average temperature and pluviometric index were obtained from "Centro Integrado de Informações Agrometeorológicas" (CIIAGRO) of "Instituto Agronômico de Campinas" (IAC). Milk samples with positive results in the microbiological test showed average SCC of 137,720 cells/mL in the dry period and 190,309 cells/mL in the rainy period. Although a higher number of isolated microorganisms was observed in buffalo milk samples during the rainy period (69/600) compared to the dry period (50/442), the season had no significant effect on the frequency of isolation of microorganisms. The main genera of microorganisms isolated were coagulase-negative Staphylococcus (38.4%), Streptococcus agalactiae (28.8%), and Bacillus spp. (7.56%) during the dry season and Corynebacterium sp. (23.5%), Streptococcus spp. (32.3%), and Streptococcus agalactiae (9.24%) during the rainy period. Multidrug resistance was observed in 30.1% of the isolated microorganisms...


O objetivo do presente estudo foi avaliar a contagem de células somáticas, a prevalência e a etiologia da mastite bubalina nas estações seca e chuvosa em um rebanho de bubalinos do município de Analândia, estado de São Paulo, Brasil. Adicionalmente, verificou-se o perfil de sensibilidade antimicrobiana dos micro-organismos isolados nas amostras de leite das búfalas. Foram avaliadas 1.042 amostras de leite de búfalas da raça Murrah pertencentes a uma propriedade rural localizada no município de Analândia-SP, obtidas no período de maio de 2011 e novembro de 2012. Após o exame físico da glândula mamária, foram realizados o teste da caneca de fundo escuro e o California Mastits Test (CMT); em seguida, foram colhidas, de forma asséptica, amostras de 50mL de leite de cada quarto mamário, para a contagem de células somáticas (CCS) em aparelho automático e exame microbiológico. Também, foi avaliado o perfil de sensibilidade antimicrobiana a ampicilina, cefoperazona, ceftiofur, enrofloxacina, gentamicina, neomicina, oxacilina, penicilina e sulfametoxazol/trimetoprim, pelo método de difusão em disco. A temperatura média e o índice pluviométrico mensais foram obtidos no Centro Integrado de Informações Agrometeorológicas (CIIAGRO) do Instituto Agronômico de Campinas (IAC). Notou-se que as amostras de leite com resultado positivo no exame microbiológico apresentaram CCS média de 137.720 células/mL, no período seco, e 190.309 células/mL, no período chuvoso. Embora tenha se constatado maior índice de isolamentos de micro-organismos nas amostras de leite obtidas no período chuvoso (69/600) do que no período seco (50/442), a frequência de isolamentos não foi influenciada significativamente pela estação do ano...


Subject(s)
Animals , Buffaloes , Mammary Glands, Animal , Mastitis, Bovine/immunology , Anti-Infective Agents/analysis , Milk Sickness/microbiology , Milk Sickness/veterinary
5.
Int. j. morphol ; 31(2): 706-715, jun. 2013. ilus
Article in English | LILACS | ID: lil-687128

ABSTRACT

The porcine transmissible gastroenteritis is a highly severe contagious disease, caused by virus of the Coronaviridae family, genus Coronavirus. Its epizootic shape can reach a rate of up to 100 percent mortality in piglets under two weeks of age as a result of severe dehydration. In this study fragments of small intestine and stool samples were collected from 75 autopsied pigs from properties. The samples of the fragments were frozen and sent to the Laboratory of Electron Microscopy, Instituto Biológico, SP, Brazil, for histological and transmission electron microscopic analyses. According to histological H&E technique, atrophy, villous necrosis and destruction of the enterocytes were observed in 35 (46.6 percent) out of the 75 fragments of the small intestine samples. On the immunohistochemistry technique 19 (25.3 percent) fragments were positively stained with DAB in the Ag-Ac reaction (MabTGEV). In 19 (25.3 percent) positive samples analyzed by in situ hybridization, a brown stain of enterocytes was observed, mainly in the epithelial cells of the villi. By the negative staining technique, we visualized enveloped, pleomorphic coronavirus particles, with typical radial projections resembling solar corona, with 140 nm diameter in 21 samples (28 percent) of the small intestine fragments and in 16 (21.3 percent) stool samples. In the ultrathin sections of 21 (28 percent) samples of small intestine, complete coronavirus particles with 80 nm diameter were seen among the microvilli and in the cytoplasm of epithelial cells. Immature particles with 60 nm diameter, budding from cell membrane and from a rough endoplasmic reticulum and also inside the vacuoles were visualized. In 19 (25.3 percent)...


La gastroenteritis transmisible porcina se caracteriza por ser una enfermedad altamente contagiosa y aguda, causada por virus de la familia Coronaviridae, género Coronavirus. Su forma epizoótica puede alcanzar una tasa de hasta 100 percent de mortalidad en lechones con menos de dos semanas de edad, como resultado de deshidratación severa. En este trabajo se recogieron 75 fragmentos de intestino delgado y muestras de heces de 75 cerdos autopsiados. Las muestras se congelaron y se enviaron al Laboratorio de Microscopía Electrónica, del Instituto Biológico, SP, Brasil, para el análisis histológico y por microscopía electrónica de transmisión. Por la técnica histológica de H&E fue observado atrofia y necrosis de la vellosidades además de la destrucción de los enterocitos en 35 (46,6 ppor ciento) de las 75 muestras de fragmentos del intestino delgado. Por inmunohistoquímica 19 (25,3 por ciento) de los fragmentos se tiñeron positivamente por el DAB en la reacción Ag-Ac (MabTGEV). En 19 (25,3 por ciento) muestras positivas mediante hibridación in situ, se observó tinción marrón de los enterocitos, principalmente en las células epiteliales de las vellosidades. Por la técnica de coloración negativa, se observaron partículas de coronavirus, encapsuladas, pleomórficas con proyecciones radiales típicas, en forma de corona solar, midiendo alrededor de 140 nm de diámetro en 21 (28 por ciento) muestras de fragmentos del intestino delgado y en 16 (21,3 por ciento muestras de heces. En cortes ultra finos de 21 (28 por ciento) fragmentos de intestino delgado fueron visualizadas partículas de coronavirus completas con 80 nm de diámetro entre las microvellosidades y en el citoplasma de las células epiteliales y partículas inmaduras de 60 nm de diámetro brotando de las membranas celulares y del retículo endoplasmático rugoso y en el interior de las vacuolas. En 19 (25,3 por ciento)...


Subject(s)
Animals , Coronavirus , Gastroenteritis, Transmissible, of Swine/pathology , Intestine, Small/pathology , Feces/virology , Immunohistochemistry , Intestine, Small/virology , Microscopy, Electron, Transmission , Swine
6.
Int. j. morphol ; 31(2): 777-784, jun. 2013. ilus
Article in English | LILACS | ID: lil-687137

ABSTRACT

Papillomaviruses, belonging to the Papillomaviridae family, are small oncogenic viruses, causing papillomas and fibropapillomas in the mucosal and cutaneous epithelia of several animals. In bovine species, thirteen types (BPV 1-13) were characterized to date. In this study, the occurrence of papillomatosis in four outbreaks in cattle herds, coming from Brazilian states were registered. The papillomatous lesions were found located in the teats, udders, head and neck. Under the transmission electron microscope, by the negative staining technique, it was possible to visualized rounded-format papillomavirus, with icosahedral symmetry, characterized as "full" and "empty" particles, measuring on average 60 nm in diameter, in all the 40 samples observed of skin lesion fragments. The ultrathin sections revealed the presence of groups of viral, intranuclear, rounded particles measuring 35 nm in diameter and tubular particles with a diameter of 35-39 nm. At immunoelectron microscopy technique, positivity obtained was marked by the presence of aggregates of viral particles formed by the antigen-antibody interaction. In the immunocytochemistry technique, the antigen-antibody reaction showed colloidal gold particles evenly distributed over the surface of the virus. These results showed the importance of the transmission electron microscopy techniques in the diagnosis of bovine papillomatosis that can be used in routine procedures to identify viral agent of this important disease.


Los virus del papiloma que pertenecen a la familia Papillomaviridae son pequeños virus oncogénicos que causan papilomas y fibropapilomas en epitelio cutáneo y mucoso de distintas especies de animales. En el ganado vacuno, trece tipos (BPV- 1-13) se caracterizaron hasta el momento. En este estudio, se documenta la ocurrencia de cuatro brotes de papilomatosis en los rebaños de ganado, procedentes de estados brasileños. Las lesiones papilomatosas se localizaron en los pezones, la ubre, la cabeza y el cuello. Al microscopio electrónico de transmisión, en la técnica de tinción negativa fueran visualizadas partículas del virus del papiloma redondeadas, con simetría icosaédrica, caracterizadas como "llenas" y "vacías", midiendo unos 60 nm de diámetro en todas las 40 muestras de fragmentos de lesión de piel estudiado. Los cortes ultra finos mostraron la presencia de grupos de partículas virales, intranucleares redondeadas con 35 nm de diámetro y tubulares 35-39 nm de diámetro. En la técnica de microscopía inmunoelectrónica, la positividad obtenida se caracterizó por la presencia de agregados de partículas virales formadas por la interacción antígeno-anticuerpo. En la aplicación de la técnica de inmunocitoquímica, la reacción antígeno-anticuerpo mostró partículas de oro coloidal distribuidos de manera uniforme sobre la superficie del virus. Estos resultados muestran la importancia de las técnicas de microscopía electrónica de transmisión en el diagnóstico de papilomatosis bovina, que pueden ser utilizados en los procedimientos de rutina para la identificación del agente viral causante de esta importante enfermedad.


Subject(s)
Humans , Animals , Cattle Diseases/pathology , Cattle Diseases/virology , Papillomaviridae/ultrastructure , Brazil , Cattle , Disease Outbreaks , Immunohistochemistry , Microscopy, Electron, Transmission
7.
Int. j. morphol ; 30(2): 723-730, jun. 2012. ilus
Article in English | LILACS | ID: lil-651859

ABSTRACT

Diseases caused by avian paramyxovirus (APMV) occur in commercial, captive and wild birds worldwide, demonstrating the significant economic and ecological importance of these agents. Paramyxoviruses belong to the paramyxoviridae family, paramyxovirinae subfamily and avulavirus genus. During the period 2000 to 2011, stool and small intestine samples of 1647 birds species were sent to the Laboratory of Electron Microscopy, Biological Institute of São Paulo, Brazil, for diagnosis of viral agents. The samples were processed by negative staining (rapid preparation) and resin embedding techniques. Under the transmission electron microscope by negative staining technique, in 294 (17.8 percent) samples of 1647 were visualized paramyxovirus particles pleomorphic, roughly spherical or filamentous, measuring 100 to 500 nm of diameter containing an envelope covered with spikes and characteristic helical herring-bone-like nucleocapsid measuring 15 to 20 nm in diameter. Ultrathin sections of the small intestine fragments revealed the presence of amorphous granular intracytoplasmic inclusions surrounded by membrane and containing viral nucleocapsid measuring 10-14 nm in diameter. Immature particles budding from cell membranes, pleomorphic, spherical and tubular particles containing viral nucleocapsid strands, and the complete particles measured up to 170 nm in diameter were seen in the cytoplasm. Intranuclear inclusions containing viral nucleocapsid were also visualized. Nuclei showed a marginalized chromatin.


Las enfermedades causadas por paramixovirus (APMV) ocurren mundialmente, tanto en aves de corral, en aquellas en vida libre o en cautiverio, lo que demuestra la importancia económica y ecológica de estos virus. El paramixovirus aviario pertenece a la familia paramyxoviridae, subfamilia paramyxovirinae y género avulavirus. Durante el periodo de 2000 a 2011, muestras de heces y fragmentos del intestino delgado de 1647 especies de aves han sido enviados al Laboratorio de Microscopía Electrónica, Instituto Biológico de São Paulo, para el diagnóstico de agentes virales. Las heces y fragmentos del intestino delgado, se procesaron por las técnicas de contraste negativo (preparación rápida) y la inclusión en resina. Al microscopio electrónico de transmisión mediante la técnica de contraste negativo se visualizaron en muestras de 294 aves, partículas de paramixovirus, pleomórficas, más o menos esféricas o filamentosas, de 100 a 500 nm de diámetro que contenían un sobre cubierto por púas que presentaban característica helicoidal, con nucleocapside tipo espiga, midiendo de 15 a 20 nm de diámetro. Secciones ultrafinas de los fragmentos del intestino delgado, revelaron en el citoplasma la presencia de inclusiones granulares amorfas rodeadas por una membrana, contiendo nucleocapside viral midiendo de 10-14 nm de diámetro, partículas inmaduras brotando de las membranas celulares, partículas virales tubulares, esféricas o pleomórficas que contenían filamentos nucleocapside. Estas partículas completas alcanzaban a los 170 nm de diámetro. Fueron observadas también, inclusiones intranucleares contiendo nucleocapside viral. Los núcleos mostraron una cromatina marginal.


Subject(s)
Animals , Birds/virology , Avulavirus Infections/diagnosis , Microscopy, Electron, Transmission , Paramyxoviridae/isolation & purification
8.
Int. j. morphol ; 30(2): 761-768, jun. 2012. ilus
Article in English | LILACS | ID: lil-651864

ABSTRACT

In this study thirty shrimp samples from commercial marine shrimp (L. vannamei) farms of southern region of Brazil were obtained. Hepatopancreas and shell scrapings fragments collected in these animals were processed by transmission electron microscopy using negative staining (rapid preparation), immunoelectron microscopy and immunocytochemistry (immunolabelling with colloidal gold particles) techniques. On the transmission electron microscopy a great number of white spot virus particles, ovoid or bacilliform-to-ellipsoid, measured 230-290 nm in length and 80-160 nm in diameter with intra-nuclear projections were visualized by the negative staining technique in 27 (90 percent) out of 30 samples examined. Using immunoelectron microscopy technique, the anti-VP 664 serum agllutinated a large number of particles formed by antigen-antibody interaction. In the immunocytochemistry technique, the antigen-antibody reaction was styrongly marked by the particles of colloidal gold over the virus. Notably, this is the first report, to our knowledge, describing use of these microscopy techniques to study Brazilian L. vannamei marine shrimp samples; moreover, this methodology also appears to be a viable complementary tool for diagnosing the presence of the white spot virus within shrimp tissues. Importantly, these are the first photoelectron micrographs of the WSSV in Brazil.


Se obtuvieron para el estudio 30 muestras de camarones marinos comerciales (L. vannamei) de las granjas de la región sur de Brasil. Fueron procesados fragmentos de hepatopáncreas y raspados internos del cefalotórax recogidos en estos animales por microscopía electrónica de transmisión con tinción negativa (preparación rápida), inmunomicroscopía y técnicas de inmunocitoquímica (inmunomarcación con partículas de oro coloidal). En la microscopía electrónica de transmisión de un gran número de partículas de virus de la mancha blanca, ovoide o elipsoidal a baciliformes, medían 230-290 nm de longitud y 80-160 nm de diámetro. En 27 (90 por ciento) de las 30 muestras examinadas intra-nuclear proyecciones se visualizaron mediante la técnica de tinción negativa. Utilizando una técnica de inmunomicroscopía electrónica, el anti-suero VP 664 reunió a un gran número de partículas formadas por la interacción antígeno-anticuerpo. En la técnica de inmunocitoquímica, la reacción antígeno-anticuerpo fue fuertemente reforzada por las partículas de oro coloidal en los virus. En particular, en Brasil este es el primer informe, a nuestro entender, que describe el uso de estas técnicas de microscopía en muestras de camarón marino L. vanamei. Además, esta metodología también parece ser una herramienta complementaria viable para diagnosticar la presencia del virus de la mancha blanca en tejidos de camarón. Es importante destacar que estas son las primeras fotos en microscopia electrónica del WSSV obtenidas en Brasil.


Subject(s)
Animals , DNA Virus Infections/pathology , Penaeidae/virology , White spot syndrome virus 1 , Brazil , Decapoda/virology , Gold Colloid , Immunohistochemistry/methods , Microscopy, Electron , Negative Staining
9.
Int. j. morphol ; 29(2): 537-542, June 2011. ilus
Article in English | LILACS | ID: lil-597488

ABSTRACT

Circovirus are viral agents that cause disease in avian species. The main clinical symptoms of the disease are immunosuppression and, in young birds feather disorders. In neonates, the disease is known as "black spot" and characterized by abdominal enlargement, hepatobiliar congestion and failure to thrive. Also, it wasobserved in adult infected birds with other symptoms and clinical signs, such as enteritis, sinusitis, rhinitis, tracheitis, bronchopneumonia, myocarditis, nephritis, splenitis, dyspnea, anorexia, depression leading to high mortality. In April 2008, 317 saffron finch were apprehended during an illegal commercialization and were forwarded to the Wild Animals Recovery Center of the Tiete Ecological Park. Subsequently, 101 (31.66 percent) died and 20 of these were selected and sent to the Laboratory of Electron Microscopy, Biological Institute of São Paulo, Brazil to investigate possible etiological agents. After necropsy of these animals fragments of lung, liver and small intestine were processed using negative staining and resin embedding techniques. On the transmission electron microscopy, the negative staining technique allowed observation in 20 (100 percent) samples of liver suspension, a great number of particles with morphology similar to the circovirus, spherical, non-enveloped, isometric, characterized as "complete" and "empty ", measuring between 17 and 20 nm in diameter. By the resin embedding technique, oval or rounded viral intracytoplasmatic inclusion bodies, containing viral particles disposed in paracrystalline arrays or loose arrangements were observed in ultrathin sections on the 20 (100 percent) samples of liver suspension. This is the first report on the presence of circovirus in saffron finch (Sicalis flaveola spp.).


Circovirus son agentes virales que causan enfermedad en las aves. Los principales síntomas clínicos de la enfermedad son la inmunosupresión y desórdenes en las plumas de aves jóvenes. En los recién nacidos la enfermedad se conoce como "punto negro" y se caracteriza por distensión abdominal, congestión hepatobiliar y falla en el desarrollo. También se han observado en las aves adultas infectadas, otros síntomas y signos clínicos, tales como la enteritis, sinusitis, rinitis, traqueitis, bronconeumonía, miocarditis, nefritis, esplenitis, disnea, anorexia, depresión llevando también a alta mortalidad. En abril de 2008, durante una comercialización ilegal, fueron confiscados 317 canarios venezolanos, siendo llevados al Centro de Rehabilitación de Animales Silvestres del Parque Ecológico de Tietê. Posteriormente, 101 (31,66 por ciento) aves murieron y entre éstas fueron seleccionadas 20 y enviadas al Laboratorio de Microscopía Electrónica, Instituto Biológico de São Paulo, para identificar el posible agente etiológico. Después de la necropsia, fragmentos de pulmón, hígado e intestino delgado se procesaron mediante las técnicas de tinción negativa y de inclusión en resina. A través de microscopía electrónica de transmisión, la técnica de tinción negativa permitió visualizar en las 20 muestras (100 por ciento) de suspensión del hígado, un gran número de partículas con morfología similar al circovirus, esférico, sin envoltura, isométrico, que se caracteriza como "completa" y "vacío", que miden entre 17 y 20 nm de diámetro. Mediante la técnica de cuerpos de inclusión en resina se observó en el 100 por ciento de las muestras de suspensión de hígado, cuerpos de inclusión virales intracitoplasmáticos, redondeadas u ovales, con partículas de virus dispuestos en matrices paracristalinas o sueltas en los cortes ultrafinos. Este es el primer reporte de la presencia de circovirus en canarios Venezolanos (Sicalis flaveola spp.).


Subject(s)
Animals , Canaries , Circovirus/ultrastructure , Liver/virology , Microscopy, Electron, Transmission
10.
Int. j. morphol ; 29(2): 628-635, June 2011. ilus
Article in English | LILACS | ID: lil-597504

ABSTRACT

The pigeon feces are vehicle of diseases both for humans and other animal species. In these birds, the most important viral diseases of the digestive tract are transmitted by the paramyxovirus, adenovirus and coronavirus. Avian paramyxoviruses have been isolated from a variety of species of free living and domestic birds worldwide, with several symptoms and clinical signs and economic losses. Paramyxoviruses belong to the Paramyxoviridae family and Avulovirus genus that includes nine serotypes (APMV 1 to 9). Avian adenoviruses belong to the Adenoviridae family and Aviadenovirus genus. In pigeons, cause classical adenovirosis and necrotizing hepatitis. The respiratory and enteric tracts are common targets of coronavirus. They belong to the Coronaviridae family and to 3a and 3c groups. In this study, we described the presence of viral agents in free-living pigeon feces (Columba livia) from the city of São Paulo, Brazil. The feces were processed by negative staining technique (rapid preparation) for transmission electron microscopy. In this technique paramyxoviruses particles, pleomorphic, roughly spherical or filamentous, measuring 100 to 500 nm of diameter containing an envelope covered by spikes, with characteristic helical herring-bone-like nucleocapsid, measuring 15 to 20 nm in diameter, were visualized in 45 (79 percent) out of 57 feces samples. In 2 (3.5 percent) samples, paramyxovirus and adenovirus particles were simultaneously visualized. Adenovirus particles were isometric, spherical, characterized as "complete "or" empty ", measuring between 70 and 90 nm in diameter. Paramyxovirus and coronavirus particles were detected in 3 (5.2 percent) samples. Coronaviruses were pleomorphic with a diameter of 75-160 nm containing a solar corona-shaped envelope, with projections of approximately 20 nm of diameter. Seven (12.3 percent) samples were negative for viral particles.


Los heces de las palomas constituyen vehículos de enfermedades importantes, tanto para el Hombre como para otras especies animales. En estas aves, las enfermedades virales más importantes del tracto digestivo son transmitidas por los paramixovirus, adenovirus y coronavirus. Paramixovirus aviario, en todo el mundo, ha sido aislado de una variedad de especies de vida libre y de aves domésticas, que causan variados síntomas y señales clínicas con pérdidas económicas. El Paramixovirus pertenece a la familia Paramyxoviridae y al género Avulavirus que incluye nueve serotipos (APMV 1 a 9). Adenovirus aviario pertenece al género de la familia Adenoviridae y género Aviadenovirus. En las palomas, causan la adenovirosis clásica y la hepatitis necrotizante. El tracto respiratorio y entérico son los albos comunes de los coronavirus. Ellos pertenecen a la familia Coronaviridae y a los grupos 3a y 3c. En este trabajo, se describe la presencia de agentes virales en las heces de palomas de vida libre (Columba livia) en la ciudad de São Paulo, Brasil. Las heces se procesaron para la técnica de microscopía electrónica de transmisión, a través de la técnica de contrastación negativa (preparación rápida). A través de esta técnica fueron visualizadas las partículas de paramixovirus, pleomórficas, más o menos esféricas o filamentosas, de 100 a 500 nm de diámetro que contiene un envoltorio cubierto con espículas, con nucleocapside con características helicoidales, midiendo de 15 a 20 nm de diámetro en 45 (79 por ciento) de 57 muestras. En 2 (3,5 por ciento) muestras, fueron observadas simultáneamente partículas de paramixovirus y de adenovirus. Las partículas de adenovirus eran isométricas, esféricas, caracterizadas como “completa " o “vacía ", midiendo entre 70 y 90 nm de diámetro. Fueron analizadas en tres muestras (5,2 por ciento) las partículas de paramixovirus y coronavirus. Los coronavirus son pleomórficos, con un diámetro de 75 a 160 nm, que contiene un capa en forma de corona solar con proyecciones de aproximadamente 20 nm de diámetro. Siete (12,3 por ciento) muestras fueron negativas para las partículas virales.


Subject(s)
Animals , Columbidae , Adenoviridae/isolation & purification , Avulavirus/isolation & purification , Coronavirus/isolation & purification , Feces/virology , Adenoviridae/ultrastructure , Avulavirus/ultrastructure , Coronavirus/ultrastructure , Microscopy, Electron, Transmission
11.
Arq. Inst. Biol. (Online) ; 78(1): 129-136, jan-mar, 2011. tab
Article in Portuguese | LILACS, VETINDEX | ID: biblio-1396452

ABSTRACT

O leite integral e a bebida láctea UAT são submetidos ao mesmo tratamento térmico, mas são produtos diferentes, já que na bebida láctea é permitida a adição de até 50% de soro lácteo. Neste estudo, objetivou-se avaliar as características microbiológicas destes produtos com relação à legislação vigente do Ministério da Agricultura, Pecuária e Abastecimento (MAPA) e, ainda, da população psicrotrófica, ao longo do período de validade. Foram analisadas 150 amostras, 75 de cada produto, a partir de cinco diferentes marcas comerciais nacionais. As 15 amostras de cada marca foram subdivididas em três lotes: início, meio e final do período de validade, cada qual com cinco amostras do mesmo lote. Verificou-se para o leite que todas as amostras de leite UAT estavam de acordo com a legislação vigente. A população de psicrotróficos aumentou em 26,6% das amostras depois que foram abertas e mantidas sob refrigeração por 48 horas. Os resultados para a bebida láctea evidenciaram que 12% das amostras estavam em desacordo com o estabelecido pela legislação vigente. A população de psicrotróficos aumentou em 36% das amostras depois que foram abertas e mantidas sob refrigeração por 48 horas. Tais resultados devem servir de alerta para a necessidade de fiscalização mais rigorosa da bebida láctea UAT, pois parte das amostras não atenderam aos padrões microbiológicos estabelecidos pela legislação.


UHT milk and dairy drinks are submitted to the same heat treatment, but they are different products, since the addition of up to 50% of whey is allowed in dairy drinks. The aim of this study was to evaluate the microbiological characteristics of these products in relation to current legislation, during the course of their shelf life. In the present study, 150 samples, 75 of each type of product, from 5 different national brands were analyzed. The 15 samples of each brand were separated into 3 lots containing 5 samples each, analyzed at the beginning, middle and end of the validity period. Upon microbiological analysis all UHT milk samples were in conformance with current legislation. After the samples were opened and chilled for 48 hours, 26.6% of them presented an increased psychrotrophic population. The dairy drink results showed that 12% of the samples were out of conformance with the current legislation. After the samples were opened and chilled for 48 hours, 36% of them presented an increased psychrotrophic population. These results should be considered as an alert in regard to the need for rigorous inspection of UHT milk and dairy drink products.


Subject(s)
Whole Foods , Milk/microbiology , Food Technology/methods , Processed Food , Bacteriological Techniques/methods
12.
J. venom. anim. toxins incl. trop. dis ; 17(3): 333-347, 2011. graf, tab
Article in English | LILACS | ID: lil-597233

ABSTRACT

In this study, we evaluated the actions of Crotalus durissus cumanensis venom (CDCmV), and its crotoxin (Crtx) fraction, on renal and vascular functions in Wistar rats. In isolated perfused kidneys, CDCmV (10 µg/mL) significantly increased the perfusion pressure (PP) from 110.7 ± 2.4 to 125.3 ± 2.8 mmHg after 30 minutes. This effect was accompanied by an increased renal vascular resistance (RVR) from 5.4 ± 0.1 to 6.2 ± 0.2 mmHg/mL.g-1.min-1. We observed decreases in urinary flow (UF) from 0.13 ± 0.01 to 0.05 ± 001 mL.g-1.min-1 and glomerular filtration rate (GFR) from 0.66 ± 0.06 to 0.18 ± 0.02 mL.g-1.min-1. Crtx did not change PP or RVR, but diminished GFR (from 0.65 ± 0.05 to 0.26 ± 003 mL.g-1.min-1) and UF (from 0.11 ± 0.008 to 0.09 ± 0.008 mL.g-1.min-1). Both CDCmV and Crtx reduced the percentage of tubular transport of sodium, chloride and potassium. The cytotoxicity of these substances against MDCK cells was tested by the MTT method: only CDCmV caused a decrease in the cell viability with an IC50 of 5.4 µg/mL. In endothelium-intact isolated aortic rings, CDCmV (0.1 to 30 µg/mL) increased the sustained phenylephrine-induced contraction to a value of 130.0 ± 6.6 percent of its corresponding control, but showed a relaxant effect in endothelium-denuded preparations. Similar results were observed in aortic rings contracted with potassium (40 mM). Crtx was ineffective in aortic ring assays. Thus, it is reasonable to suggest that the renal effects induced by the CDCmV may be due to its influence on the endothelium's ability to release factors that can alter the contractile behavior of vascular smooth muscle. In conclusion, CDCmV is toxic to kidney cells. It changes parameters of the renal function including the glomerular filtration rate, renal vascular resistance and tubular transport. The actions induced by CDCmV also involve endothelium-dependent vasoactive properties. Their effects may be only partially attributed to Crtx.


Subject(s)
Animals , Female , Rats , Crotalus , Crotoxin , Rats, Wistar , Crotalid Venoms/toxicity
13.
Int. j. morphol ; 28(2): 549-555, June 2010. ilus
Article in English | LILACS | ID: lil-577152

ABSTRACT

Coronaviruses are enveloped, positive-stranded RNA, belongs to the Nidovirales order, Coronaviridae family and are classified into three groups. Groups 3a and 3c comprising the avian coronaviruses. They are worldwide in distribution and can cause respiratory, enteric and in some cases hepatitis and neurological diseases in a wide variety of animals, resulting in economic losses to breeders and breeding. In this study, we describe an outbreak of diarrhea in a herd of capybaras found in the city of Piracicaba, São Paulo, Brazil. The animals had sudden symptoms of anorexia, profuse and watery diarrhea, yellowish or greenish, dehydration, weight loss, sharp and twisting of limbs leading to death in less than a week, mostly weaned and with an age of three months. A total of 59 stool samples and 10 fragments of small intestine were processed for negative staining technique (rapid preparation). By transmission electron microscope a large number of particles with morphology similar to coronavirus, pleomorphic, rounded or elongated features with radial projections form a corona and measuring 80-140 nm in diameter, was visualized in all samples analyzed. The technique was extremely useful for rapid viral diagnosis in the affected animals.


Coronavirus están envueltos en una cadena positiva de ARN, (orden Nidovirales, familia Coronaviridae) y se clasifican en tres grupos. Grupos que integran la tercera 3c y los coronavirus aviario. Se distribuyen por todo el mundo y pueden causar enfermedades respiratorias, entéricas y, en algunos casos, hepatitis y enfermedades neurológicas en una amplia variedad de animales, resultando en pérdidas económicas a la ganadería y los criadores. En este trabajo se describe un brote de diarrea en un rebaño de capibaras en la ciudad de Piracicaba, São Paulo, Brasil. Los animales presentaron súbitamente, los síntomas de anorexia, diarrea, profusa y acuosa, de color amarillento o verdoso, deshidratación, pérdida de peso, agudo, torsión de los miembros y la muerte en menos de una semana, en su mayoría con tres meses de edad. Un total de 59 muestras de heces y 10 trozos de intestino delgado se procesaron por la técnica de contraste negativo (preparación rápida). En todas las muestras analizadas, al microscopio electrónico de transmisión se visualizó un gran número de partículas con morfología similar a los coronavirus, pleomórficas, con características redondeadas o alargadas con proyecciones radiales en forma de corona midiendo de 80-140 nm de diámetro. La técnica es sumamente útil para el diagnóstico rápido de virus en animales afectados.


Subject(s)
Animals , Coronavirus/ultrastructure , Rodent Diseases/virology , Microscopy, Electron, Transmission , Rodentia , Brazil , Coronavirus/isolation & purification , Feces/virology , Intestine, Small/virology , Staining and Labeling
14.
Int. j. morphol ; 28(2): 627-636, June 2010. ilus
Article in English | LILACS | ID: lil-577164

ABSTRACT

The barn-owl (Tyto Alba) and striped-owl (Rhinoptynx clamator) belong respectively to the families Tytonidae and Strigidae. Avian paramyxoviruses have been isolated from a variety of species of wild and domestic birds wordlwide causing diverse clinical symptoms and signs. Paramyxoviruses belong to the family Paramyxoviridae and Avulovirus genus, including nine serotypes (APMV 1 to 9). The lymphoid leukosis is a retrovirus-induced neoplasia. The avian retroviruses belong to the Retroviridae family and to the Alpharetrovirus genus. Coronaviruses can cause respiratory and enteric disease in several species of birds. They belong to the Coronaviridae family and to the groups 3a e 3c. In this study, we describe the presence of viruses in four owls, two barn owls (Tyto alba) and two striped owls (Rhinoptynx clamator), rescued from tree-lined streets of Sao Paulo, Brazil and sent to the Recovery Center of Wild Animals of the Tietê Ecological Park, where the animals died. Fragments of lung, liver and small intestine of these birds were processed for transmission electron microscopy utilizing negative staining (rapid preparation), immunoelectron microscopy and immunocitochemistry techniques. Under the transmission electron microscopy paramyxovirus particles, pleomorphic, roughly spherical or filamentous, measuring 100 to 500 nm of diameter containing an envelope covered by spikes, an herring-bone helical nucleocapsid-like structure, measuring 15 to 20 nm in diameter, were visualized in the samples of lung, liver and small intestine of all owls. In small intestine samples of the two striped-owl (owls 3 and 4) it was detected pleomorphic coronavirus particles with a diameter of 75-160 nm containing a solar corona-shaped envelope, with projections of approximately 20 nm of diameter. In liver fragments of one striped-owl (owl 4) pleomorphic particles of retrovirus with a diameter of 80-145 nm containing an envelope with short projections and diameter of 9 nm were....


La lechuza (Tyto Alba) y el búho de orejas (Rhinoptynx clamator) pertenecen respectivamente a las familias Strigidae y Tytonidae. El paramixovirus aviario se ha aislado de especies de vida silveste como las aves domésticas por todo el mundo, causando diversos síntomas clínicos. El paramixovirus pertenece a la familia Paramyxoviridae y al Avulovirus genus que incluye nueve serotipos (APMV 1 a 9). La leucosis linfoide es una neoplasia inducida por retrovirus. Los retrovirus aviarios pertenecen a la familia Retroviridae y el género Alpharetrovirus. Los coronavirus pueden causar enfermedades respiratorias y entéricas en varias especies de aves. Ellos pertenecen a la familia Coronaviridae y a los grupos 3a y 3c. En este estudio, se describe la presencia del virus en cuatro búhos, dos lechuzas (Tyto alba) y dos búhos de orejas (Rhinoptynx clamator), rescatados de las calles arboladas de São Paulo, Brasil y enviados al Centro de Recuperación de Animales Silvestres del Parque Ecológico de Tietê, donde hubo murieron los animales. Fragmentos de pulmón, delhígado y del intestino delgado de estas aves fueron procesados para microscopía electrónica de transmisión utilizando tinción negativa (preparación rápida), inmunomicroscopía y técnicas de inmunocitoquímica. Bajo microscopía electrónica de transmisión, partículas de paramixovirus, pleomórficas, aproximadamente esféricas o filamentosas, de 100 a 500 nm de diámetro con un sobre cubierto por espigas, y nucleocápside helicoidal con características de espiga, midiendo 15 a 20 nm de diámetro, fueron visualizadas en las muestras de pulmón, hígado e intestino delgado de todos los búhos. En muestras de intestino delgado de dos búho de orejas (búhos 3 y 4) se detectaron partículas pleomórficas con coronavirus de un diámetro de 75-160 nm con un sobre con forma de corona solar, con proyecciones de aproximadamente 20 nm de diámetro. En el hígado de un búho de orejas (búho 4) se observaron partículas pleomórficas de retrovirus con ...


Subject(s)
Animals , Strigiformes/anatomy & histology , Strigiformes/virology , RNA Viruses/immunology , RNA Viruses/ultrastructure , Brazil , Coronavirus/immunology , Coronavirus/ultrastructure , Immunohistochemistry , Microscopy, Electron, Transmission , Paramyxoviridae/immunology , Paramyxoviridae/ultrastructure , Retroviridae/immunology , Retroviridae/ultrastructure
15.
J. venom. anim. toxins incl. trop. dis ; 16(3): 493-504, 2010. ilus, graf, tab
Article in English | LILACS | ID: lil-557178

ABSTRACT

Snake venom proteins from the C-type lectin family have very distinct biological activities despite their highly conserved primary structure, which is homologous to the carbohydrate recognition region of true C-type lectins. We purified a lectin-like protein (BmLec) from Bothrops moojeni venom and investigated its effect on platelet aggregation, insulin secretion, antibacterial activity, and isolated kidney cells. The BmLec was purified using two chromatographic steps: affinity chromatography and reverse phase high performance liquid chromatography (HPLC). BmLec showed a dose-dependent platelet aggregation and significantly decreased the bacterial growth rate in approximately 15 percent. During scanning electron microscopy, the profile of Xanthomonas axonopodis pv. passiflorae treated with lectin disclosed a high vesiculation and membrane rupture. BmLec induced a strong and significant increase in insulin secretion at 2.8 and 16.7 mM glucose concentrations, and this effect was seen in the presence of EGTA in both experiments. BmLec (10 µg/mL) increased the perfusion pressure, renal vascular resistance and urinary flow. The glomerular filtration rate and percentages of sodium, potassium and chloride tubular transport were reduced at 60 minutes of perfusion. Renal alterations caused by BmLec were completely inhibited by indomethacin in all evaluated parameters. In conclusion, the C-type lectin isolated from Bothrops moojeni affected platelet aggregation, insulin secretion, antibacterial activity and isolated kidney function.


Subject(s)
Animals , Bothrops , Crotalid Venoms , Insulin , Kidney , Lectins, C-Type/isolation & purification , Platelet Aggregation , Chromatography, High Pressure Liquid/methods
16.
J. venom. anim. toxins incl. trop. dis ; 16(4): 614-622, 2010. ilus, graf
Article in English | LILACS | ID: lil-566161

ABSTRACT

Some snake venom proteins present enzymatic activities, such as L-amino acid oxidase (LAAO). The aim of this paper was to investigate the effect of Bothrops leucurus total venom (BleuTV) and its fraction LAAO (BleuLAAO) on bacteria, yeast, and promastigote forms of Leishmania amazonensis and Leishmania chagasi, and epimastigote forms of Trypanosoma cruzi. BleuTV was isolated with a Protein Pack 5PW® (Waters Corporation, USA), and several fractions were obtained. BleuLAAO was purified to high molecular homogeneity, and its N-terminal amino acid sequence shared a high degree of amino acid conservation with other LAAOs. BleuTV inhibited Staphylococcus aureus growth in a dose-dependent manner, with a minimum inhibitory concentration (MIC) of 25 ìg/mL, which corresponded to its minimum lethal concentration (MLC). BleuTV also inhibited the growth of promastigote forms of L. chagasi and L. amazonensis, with respective IC50 values of 1.94 ìg/mL and 5.49 ìg/mL. Furthermore, it repressed T. cruzi growth with an IC50 of 1.14 ìg/mL. However, BleuLAAO did not inhibit the growth of the microorganisms studied and was not toxic to macrophages. BleuTV had low toxicity against macrophages at the concentrations studied. In conclusion, whole venom from Bothrops leucurus inhibited the growth of some microorganisms, including S. aureus, Leishmania sp., and T. cruzi.


Subject(s)
Animals , L-Amino Acid Oxidase , Crotalid Venoms/antagonists & inhibitors , Crotalid Venoms/isolation & purification , Leishmania/microbiology , Staphylococcus aureus , Trypanosoma cruzi/microbiology
17.
Int. j. morphol ; 27(2): 543-552, June 2009. ilus
Article in English | LILACS | ID: lil-563109

ABSTRACT

The myxomatosis is a contagious worldwide disease caused by poxvirus which infects domestic and wild rabbits. In the present study we present two distinct outbreaks of myxomatosis when raising rabbits, one for commercial purpose of production of meat and skins and, another one for the commercialization of ornamental rabbits. The observed signs were ocular, auricular, nasal, testis lesions and many times scattered throughout the body of the animals. The lesions were characterized by formation of nodules that by palpation disclosed gummy or gelatinous aspect. At the transmission electron microscopy, all the skin and crust samples were analyzed by negative staining technique. A great number of particles with morphology similar to the poxvirus, some enveloped in a brick-shaped and irregular disposition of tubules on the external membrane, measuring 300x240 nm on the average were visualized. Ultra thin sections revealed the presence of intracytoplasmic inclusion bodies surrounded by membrane containing oval particles, measuring 270 x 130 nm, containing nucleus or an internal biconcave (dumbbell-shaped) core. Immature particles (empty), surrounded by membrane were also observed. In addition, intracytoplasmic electron dense inclusion bodies containing viral particles budding of dense amorphous material and intranuclear fibrillar or "digital" inclusions showing a regular striation and arranged in groups were found in the middle of granular material. The nuclei were deformed with densely condensed chromatin forming amorphous and electron dense inclusion bodies. In the immunocytochemistry technique, the antigen-antibody reaction was strongly marked by the particles of colloidal gold, emphasizing the viral particles. The techniques used in this study were important in the diagnosis of the affected animals.


La mixomatosis es una enfermedad contagiosa de distribución mundial, causada por poxvirus que infecta conejos domésticos y salvajes. En este estudio presentamos dos distintos surtos por mixomatosis que ocurrieron en producciones de conejos, una para fines comerciales de producción de carne y pieles y otra para el comercio de conejos domésticos. Las señales observadas fueron afecciones oculares, nasales, testiculares y, a veces, también distribuida por todo el cuerpo de los animales. Estas se caracterizaban por formación de nódulos que a la palpación tenían un aspecto gelatinoso o gomoso. En la microscopía electrónica de transmisión, por la técnica de contrastación negativa, se pudo observar en todas las muestras examinadas de piel y de costras, un gran número de partículas típicas de poxvirus, con envoltura y forma de ladrillo, mostrando disposición irregular de los túbulos sobre la membrana externa, midiendo 300 x 240 nm en el promedio. Cortes ultrafinos de fragmentos de piel y de costras revelaron la presencia de cuerpos de inclusión intracitoplasmáticas, envueltos por membrana y conteniendo partículas ovales, midiendo 270 x 130 nm, conteniendo núcleo o centro interno bicóncavo (forma de mancuernas). Partículas inmaduras (vacías) envueltas por membrana fueron observadas. También fueron analizados cuerpos de inclusión intracitoplasmáticos, electrodensos, conteniendo partículas virales brotando del material denso y amorfo. Fueron observadas inclusiones intranucleares fibrilares o "digitales" mostrando una estriación periódica y disposición en grupos en medio del material granular. Los núcleos estaban deformados con cromatina densamente condensada formando cuerpos de inclusiones electrodensas y amorfas. En la técnica de imunocitoquímica la reacción antígeno-anticuerpo fue intensamente marcada por las partículas de oro coloidal realzando fuertemente las partículas virales.


Subject(s)
Male , Adult , Animals , Rabbits , Myxomatosis, Infectious/complications , Myxomatosis, Infectious/prevention & control , Myxomatosis, Infectious , Myxomatosis, Infectious/blood , Myxoma virus/isolation & purification , Myxoma virus/classification , Myxoma virus/ultrastructure , Brazil/epidemiology , Microscopy, Electron, Transmission/methods , Microscopy, Electron, Transmission/veterinary , Panophthalmitis/etiology , Panophthalmitis/veterinary , Rhinitis/etiology , Rhinitis/veterinary , Testis/injuries
18.
Int. j. morphol ; 27(2): 577-585, June 2009. ilus
Article in English | LILACS | ID: lil-563113

ABSTRACT

Avianpox is a highly contagious disease infecting both commercial and wild birds, causing great damages to breeders and breeding. Caused by DNA viruses of the family Poxviridae, genus Avipoxvirus, if manifest through 3 forms, cutaneous, diphtheric and septicemic. In June 2003, during illegal commercialization of Brazilian birds, 800 wild birds (Paroaria dominicana, Sporophila caerulescens and Sporophila albogularis) were apprehended and being forwarded to the CRAS (Wild Animals Recovery Center), Tietê Ecological Park. After one month, birds presented cutaneous lesions in the beak and feet and anorexia, emaciation, locomotion difficulties, diarrhea, dehydration and death. Among the 800 birds, 500 died and 40 these (15 Paroaria dominicana, 15 Sporophila caerulescens and 10 Sporophila albogularis) were sent to the Electron Microscopy Laboratory of the Biology Institute of São Paulo, SP, to investigate viral agents. Scabs and fragments of skin lesions collected of theses birds were processed for transmission electron microscopy utilizing negative staining (rapid preparation), resin embedding and immunocitochemistry techniques. Under the transmission electron microscopy in all the analyzed samples it was visualized two types of poxvirus particles, M form, with regular spaced thread-like ridges comprising the exposed surface, measuring 280 x 230 nm; C form or stain-penetrated particle showing the dumbbell-shaped core surrounded by the outer envelope, measuring 360 x 330 nm. In the ultrathin sections obtained, three types of intracytoplasmic inclusion bodies were encountered: type A or Bollinger body, outlined by membrane, containing in its interior a great number of mature particles, measuring 200 x 300 nm, revealing the inner dumbbell-shaped core, two lateral bodies and an external envelope. In the type B electron dense inclusions bodies, viral particles budding of dense amorphous material were observed. Fibrillar inclusions constituted by ...


Avianpox es una enfermedad altamente contagiosa que infecta tanto aves comerciales como aves silvestres, causando grandes daños a lo ganadería y a la crianza. Causados por los virus de ADN de la familia Poxviridae, género Avipoxvirus, se manifiestan a través de 3 formas, cutánea, diftérica y septicémica. En junio del 2003, durante la comercialización ilegal de aves brasileñas, 800 aves silvestres (Paroaria dominicana; Sporophila caerulescens; Sporophila albogularis) fueron capturadas y remitidas al CRAS (Centro de Recuperación de Animales Silvestres), Parque Ecológico de Tietê. Después de un mes, las aves presentaron lesiones cutáneas en el pico y las patas, anorexia, adelgazamiento, dificultades de locomoción, diarrea, deshidratación y muerte. 500 de las 800 aves murieron, 40 de éstas (15 Paroaria dominicana, 15 Sporophila caerulescens y 10 Sporophila albogularis) fueron enviadas al Laboratorio de Microscopía Electrónica del Instituto de Biología de São Paulo, SP, para investigar agentes virales. Costras y fragmentos de lesiones en la piel de estas aves fueron recolectadas y se procesaron para microscopía electrónica de transmisión utilizando tinción negativa (preparación rápida), incrustación de resina y técnica inmunocitoquímica. Bajo la microscopía electrónica de transmisión en todas las muestras analizadas se encontraron dos tipos de partículas de poxvirus, Forma M, con espacios regulares como hilo que abarcan toda la superficie expuesta, midiendo 280 x 230 nm; Forma C o partícula teñida por penetración mostrando núcleo en forma de pesa rodeado por una envoltura externa, que medía 360 x 330 nm. Se encontraron en secciones ultrafinas, tres tipos de cuerpos de inclusión intracitoplásmica: el tipo A o cuerpo de Bollinger, esbozado por la membrana, que contiene en su interior un gran número de partículas maduras, que miden 200 x 300 nm, revelando núcleo interno en forma de pesa, dos cuerpos laterales y una envoltura externa. En el tipo ...


Subject(s)
Male , Animals , Femur Head/anatomy & histology , Femur Head/innervation , Femur Head/injuries , Femur Head/ultrastructure , Chickens/anatomy & histology , Chickens/growth & development , Cartilage/anatomy & histology , Cartilage/injuries , Femur Head Necrosis/veterinary
19.
J. venom. anim. toxins incl. trop. dis ; 15(1): 125-135, 2009. ilus, graf
Article in English | LILACS | ID: lil-508235

ABSTRACT

Thalassophryne nattereri (niquim) is a venomous fish responsible for numerous accidents involving fishermen in northern and northeastern Brazil. The aim of the present investigation was to evaluate the action of antivenom on renal effects caused by Thalassophryne nattereri venom. Isolated kidneys of Wistar rats were perfused with a previously dialyzed Krebs-Henseleit solution containing 6 g% bovine serum albumin. The antivenom action was studied through perfusion pressure (PP), renal vascular resistance (RVR), urinary flow (UF) and glomerular filtration rate (GFR). The niquim venom (1 miug/mL), the antivenom alone (1 miug/mL) or the venom incubated with antivenom were added to the system 30 minutes after the beginning of each perfusion. Previous works have shown venom induced-alterations of renal function parameters. In the isolated rat Kidney, T. nattereri venom (1 miug/mL) increased the perfusion pressure and renal vascular resistance at 60, 90 and 120 minutes. UF and GFR also increased at 60, 90 and 120 minutes when compared with the control group; however, no effects were observed on the percent of sodium (% TNa more control equal 81.1 more or less 0.86; % TNa more 60 equal 78.04 more or less 1.18; % TNa more 90 equal -5.16 more or less 3.34; %TNa more 120 equal 79.49 more or less 0.87) and potassium (%TKcontrol equal 72.29 more or less 1.12; %TK more 60 equal 75.41 more or less 0.65; % TK more 90 equal 71.23 more or less 2.55; % TK more 120 equal 76.62 more or less 1.04) tubular transporto. The administration of the antivenom (1 miug/mL) incubated with venom (1 miug/mL) reduced the changes in PP, RVR, UF and GFR provoked by Thalassophryne nattereri venom. The group perfused with venom alone showed a moderate deposit of a proteinaceous material in the tubules and urinary space.(...)


Subject(s)
Animals , Male , Rats , Antivenins , Kidney/anatomy & histology , Kidney/pathology , Fish Venoms/antagonists & inhibitors , Fish Venoms/toxicity
20.
Arq. bras. med. vet. zootec ; 59(6): 1563-1569, dez. 2007. tab
Article in Portuguese | LILACS | ID: lil-476132

ABSTRACT

Pesquisaram-se a presença de Bacillus cereus e a produção de enterotoxinas produzidas por esses microrganismos em 120 amostras de diversos tipos de leite. Bacillus cereus foi isolado e identificado em 22 (73,3 por cento), 15 (50,0 por cento), 29 (96,7 por cento) e quatro (13,3 por cento) amostras de leite em pó, cru, pasteurizado e UAT (longa vida), respectivamente. Para a detecção de enterotoxinas pela técnica da alça ligada de coelho, foram positivos, respectivamente, três (13,6 por cento), um (7,1 por cento) e 10 (35,7 por cento) microrganismos isolados das amostras de leite em pó, leite cru e leite pasteurizado. Pelo teste de aumento de permeabilidade vascular, dois (9,1 por cento), um (7,1 por cento), um (3,6 por cento) e um (4,0 por cento) microrganismos isolados de leite em pó, cru, pasteurizado e UAT apresentaram-se enterotoxigênicos, respectivamente. O uso da técnica de aglutinação passiva em látex demonstrou a produção da toxina diarréica por três (33,3 por cento), sete (63,6 por cento), quatro (30,8 por cento) e oito (80,0 por cento) microrganismos isolados, respectivamente, de leite em pó, cru, pasteurizado e UAT. Os resultados indicam um risco potencial, podendo colocar em risco a saúde dos consumidores desses produtos


A hundred and twenty samples of different types of milk were examined to the presence of Bacillus cereus and the enterotoxigenic potential of the isolated strains. Bacillus cereus was isolated and identified in 22 (73.0 percent), 15 (50.0 percent), 29 (96.7 percent) and four (13.3 percent) samples of powder, raw, pasteurized and UHT milk, respectively. The enterotoxigenicity detection using the rabbit ileal loop assay showed positive, respectively, three (13.6 percent), one (7.1 percent) and 10 (35.7 percent) isolated strains from powder, raw and pasteurized milk. Using vascular permeability activity assay two (9.1 percent), one (7.1 percent), one (3.6 percent) and one (4.0 percent) isolated strains from powder, raw, pasteurized and UHT milk were positive, respectively. The reversed passive latex agglutination test showed diarrheal toxin production by three (33.3 percent), seven (63.6 percent), four (30.8 percent) and eight (80.0 percent) strains isolated from, respectively, powder, raw, pasteurized and UHT milk. Those results indicated a potential hazard and may put in risk the health of the consumers of these products


Subject(s)
Bacillus cereus/pathogenicity , Enterotoxins , Milk/microbiology , Epidemiology
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